摘要
目的观察外源性转化生长因子β1(TGF-β1)对原代急性早幼粒细胞白血病细胞(APL)凋亡的影响,并探讨其可能机制。方法将APL细胞分为4组,对照组不加任何药物,实验组用终浓度为1、5、10 ng/mL TGF-β1处理(实验1、2、3组),分别处理24、48、72 h,采用瑞氏—吉姆萨染色法检测细胞凋亡,流式细胞仪检测细胞周期,免疫组化法检测TGF-β1、P27Kip1、Cyclin E、bcl-2蛋白,RT-PCR技术检测TGF-β1、P27Kip1、Cyclin E、bcl-2 mRNA。结果与对照组比较,实验1、2、3组24、48 h细胞凋亡率升高(P均<0.05);与同组24 h比较,实验1、2、3组48 h细胞凋亡率升高(P均<0.05)。对照组细胞培养48 h未见明显细胞周期阻滞;与对照组比较,实验1组48 h细胞周期阻滞于G1期,实验2、3组APL细胞周期明显阻滞于G1期。与对照组比较,实验2组TGF-β1、P27Kip1蛋白水平升高,Cyclin E、bcl-2蛋白降低(P均<0.05)。1'、2'β-actin条带亮度相同,实验2组(2道)P27Kip1mRNA条带亮度与对照组(1道)相同,P27Kip1mRNA水平较对照组变化不明显;1'、2'β-actin条带亮度相同,实验2组(2道)条带亮度明显高于对照组(1道);实验2组Cyclin E条带与对照组比较亮度减低,Cyclin E表达下调。结论外源性TGF-β1通过上调TGF-β1、P27Kip1及下调Cyclin E、bcl-2作用,诱导细胞发生凋亡,使细胞阻滞于G1期。
Objective To observe the effect of exogenetic TGF-β1 on apoptosis of primary acute promyelocytic leukae- mia （APL） cells and to investigate the mechanism. Methods APL cells were divided into 4 groups： the control group, the experimental groups 1, 2 and 3. The control group was not treated with any drugs, but the experimental groups 1, 2 and 3 were separately treated with TGF-β1 at the concentrations of 1,5, and 10 ng/mL for 24, 48 and 72 h. The apoptosis was de- tected by Swiss-Jim staining method and cell cycle was detected by flow cytometry. The expression of TGF-β1, P27Kip1 , Cyc- lin E and bcl-2 protein and mRNA in APL cells was detected by immunohistochemistry and RT-PCR. Results Compared with the control group, the apoptosis rate of the experimental groups 1, 2 and 3 at 24 h and 48 h was increased （ all P 〈 0.05 ） ; in each group except the control group, the apoptosis rate at 48 h was higher than that at 24 h （ all P 〈 0.05 ）. In the control group, after being cultured for 48 h, no cell cycle arrest was shown ; compared with the control group, after being cul- tured for 48 h, the cell cycle of APL cells in the experimental group 1 was blocked in G1 stage and the cell cycle of APL cells in the experimental groups 2 and 3 was significantly blocked in G1 stage. Compared with the control group, the expression lev- els of TGF-β1 and P27Kipl proteins were increased while Cyclin E, bcl-2 levels were decreased, in APL cells （P 〈 0.05）. 1＇, 2＇ 13-actin band had the same brightness, and the P27Ki~ mRNA band was the same in the experimental group 2 （2＇） and the control group（ 1＇）, P27Kipl mRNA had no significant changes as compared with the control group; the brightness of Cyclin E band was reduced in the experimental group 2 as compared with that of the control group and the expression of Cyclin E was down-regulated. Conclusion Exogenetic TGF-β1 can induce apoptosis and block the cell cycle in the G1 stage by up-regula-ting the TGF-β1 and P27Kipl, and down-regulating Cyclin E and bcl-2.
出处
《山东医药》
CAS
2014年第11期22-25,共4页
Shandong Medical Journal
基金
沈阳市科学技术项目计划(F12-277-1-21)
作者简介
王蕾（1981-），女，讲师，研究方向为恶性血液病发病机制。E-mail：leiwen000@sina．com
通信作者：李艳（1958-），女，教授，研究方向为恶性血液病发病机制。E-mail：liyan2@medmail．com．cn
